Heat stress induced enhancement of heat shock protein gene activity in the honey bee (Apis mellifera)

Summary We employed in vitro translation of mRNA and product separation using SDS-PAGE to examine the heat-shock response of the worker honey bee. Increases in the levels of 6 translatable RNA populations were observed following heat stress. The greatest response was observed among bees aged 9 days. Slight levels of induction of 70 and 82 kDa heat shock proteins were evident among bees taken directly from the colony.     

Heat shock response in Drosophila.

Major alterations in genetic activity have been observed in every organism after exposure to abnormally high temperatures. This phenomenon, called the heat shock response, was discovered in the fruit fly Drosophila. Studies with this organism led to the discovery of the heat shock proteins, whose genes were among the first eukaryotic genes to be cloned. Several of the most important aspects of the regulation of the heat shock response and of the functions of the heat shock proteins have been unraveled in Drosophila.     

Heat shock response inDrosophila

Major alterations in genetic activity have been observed in every organism after exposure to abnormally high temperatures. This phenomenon, called the heat shock response, was discovered in the fruit flyDrosophila. Studies with this organism led to the discovery of the heat shock proteins, whose genes were among the first eukaryotic genes to be cloned. Several of the most important aspects of the regulation of the heat shock response and of the functions of the heat shock proteins have been unraveled inDrosophila.     

Acute heat/exercise stress in rats: effects on fluid and electrolyte regulatory hormones

Adult, male rats were exercised in the heat until hyperthermic exhaustion ensued. Plasma aldosterone levels were significantly (p less than 0.001) elevated after 8 min of exercise and remained increased throughout the exercise and recovery periods. Alternatively, plasma angiotensin I levels were unaffected during exercise, but increased significantly (p less than 0.001) during the recovery period. These rapid elevations in hormonal levels may be part of a sympathicoadrenal response to heat/exercise stress as well as an adaptational response to maintain plasma volume during and following exercise in the heat.     

Heat production by balling in the Japanese honeybee,Apis cerana japonica as a defensive behavior against the hornet,Vespa simillima xanthoptera (Hymenoptera: Vespidae)

Summary As an effective counterattack strategy against predacious hornets, especiallyVespa simillima xanthoptera, workers ofApis cerana japonica showed a distinct balling reaction, usually involving 180–300 bees. This produced heat for as long as 20 min, giving rise to temperatures inside the ball higher than 46°C, which is lethal to the hornet but not to the bees.     

The emerging role for sphingolipids in the eukaryotic heat shock response

Eukaryotic cells have a highly conserved response to an increase in temperature, termed the heat shock response. Recent research has revealed multiple roles for various sphingolipids in the heat shock responses of both yeast and mammalian cells. Heat stressed or shocked yeast and mammalian cells have an acute activation of serine palmitoyltransferase, resulting in the de novo biosynthesis of sphingolipids. Also, both mammalian and yeast cells were shown to increase ceramide levels upon heat stress or shock. In yeast cells, several functions have emerged for the de novo produced sphingoid bases in terms of the heat stress response. These functions include a role in accumulation of trehalose, a role in the heat-induced transient G0/G1 cell cycle arrest and phytosphingosine activation of a ubiquitin protein degradation pathway. However, in mammalian systems, ceramides have been demonstrated as bioactive lipids. Ceramides produced in response to heat shock were demonstrated to induce the production of c-jun, leading to apoptosis, and to be upstream of dephosphorylation of serine-rich proteins. Increasingly, sphingolipids are emerging as bioactive signaling molecules involved in numerous aspects of the eukaryotic heat shock response.     

Genetic structure and division of labor in honeybee societies

Summary Recent studies have demonstrated a genotypic component to the division of labor among worker honeybees. However, these studies used artificially-selected strains of bees or colonies derived from queens that were instrumentally inseminated with the semen from very few males. We present evidence for genotypic variability among groups of workers performing tasks in colonies with naturally-mated queens. These results demonstrate that genetic structure is a level of social organization in honeybees.     

Altered proteostasis in aging and heat shock response in C. elegans revealed by analysis of the global and de novo synthesized proteome

Protein misfolding and aggregation as a consequence of impaired protein homeostasis (proteostasis) not only characterizes numerous age-related diseases but also the aging process itself. Functionally related to the aging process are, among others, ribosomal proteins, suggesting an intimate link between proteostasis and aging. We determined by iTRAQ quantitative proteomic analysis in C. elegans how the proteome changes with age and in response to heat shock. Levels of ribosomal proteins and mitochondrial chaperones were decreased in aged animals, supporting the notion that proteostasis is altered during aging. Mitochondrial enzymes of the tricarboxylic acid cycle and the electron transport chain were also reduced, consistent with an age-associated energy impairment. Moreover, we observed an age-associated decline in the heat shock response. In order to determine how protein synthesis is altered in aging and in response to heat shock, we complemented our global analysis by determining the de novo proteome. For that, we established a novel method that enables both the visualization and identification of de novo synthesized proteins, by incorporating the non-canonical methionine analogue, azidohomoalanine (AHA), into the nascent polypeptides, followed by reacting the azide group of AHA by ‘click chemistry’ with an alkyne-labeled tag. Our analysis of AHA-tagged peptides demonstrated that the decreased abundance of, for example, ribosomal proteins in aged animals is not solely due to degradation but also reflects a relative decrease in their synthesis. Interestingly, although the net rate of protein synthesis is reduced in aged animals, our analyses indicate that the synthesis of certain proteins such as the vitellogenins increases with age.     

Microbial thermosensors

Temperature is among the most important of the parameters that free-living microbes monitor. Microbial physiology needs to be readjusted in response to sudden temperature changes. When the ambient temperature rises or drops to potentially harmful levels, cells mount protective stress responses—so-called heat or cold shock responses, respectively. Pathogenic microorganisms often respond to a temperature of around 37°C by inducing virulence gene expression. There are two main ways in which temperature can be measured. Often, the consequences of a sudden temperature shift are detected. Such indirect signals are known to be the accumulation of denatured proteins (heat shock) or stalled ribosomes (cold shock). However, this article focuses solely on direct thermosensors. Since the conformation of virtually every biomolecule is susceptible to temperature changes, primary sensors include DNA, RNA, proteins and lipids.     

Zum phototaktischen Laufverhalten junger Honigbienen (Apis mellifica L.)

Summary The number of phototactic runs towards low light intensities of young bees (Apis mellifica L.) depends on their age. On the 1st day there is almost no response. After the 4th day a light intensity as low as 0.1 Lux induces a phototactic run. This increase of response coincides with the performance of the 1st exercise flight.     

Interspecific diversity and uniformity of flower colour patterns as cues for learned discrimination and innate detection of flowers

The colour patterns of angiosperm flowers visited by bees and hoverflies were shown to provide a dual signalling system, giving the flower-visitors visual orientation cues for the initial detection of flowers, as well as for learned discrimination. Pollinators show an innate response to interspecifically uniform colour signals, which enable them to detect flowers and to orient towards the site of reward. Interspecifically diverse colour signals provide pollinators with cues for learned discrimination between flowers of different species.     

Polymorphisms in mitochondrial DNA of European and Africanized honeybees (Apis mellifera)

This study demonstrates polymorphisms in both the length and in the restriction enzyme cleavage sites of honeybee mitochondrial DNA (mtDNA). The levels of variation are typical of those found in other metazoan species. These polymorphisms are potentially useful for the identification of Africanized bees in the western hemisphere and for study of honeybee phylogenetics.     

Polymorphisms in mitochondrial DNA of european and Africanized honeybees (Apis mellifera)

Summary This study demonstrates polymorphisms in both the length and in the restriction enzyme cleavage sites of honeybee mitochondrial DNA (mtDNA). The levels of variation are typical of those found in other metazoan species. These polymorphisms are potentially useful for the identification of Africanized bees in the western hemisphere and for study of honeybee phylogenetics.     

Genotypic variation in foraging responses to environmental stimuli by honey bees,Apis mellifera

We experimentally tested a model predicting that colony-level genotypic diversity contributes to colonylevel foraging flexibility in honey bees. We established a colony into which we placed individually marked workers from three genetically distinct groups. The colony was placed in an enclosure that contained feeding stations with pollen and sugar syrup. Foraging resources, stores of pollen and the quantity of brood within the colony were varied temporally. Individual foragers switched between resource types in response to changes in relative resource quality and colony need, demonstrating flexibility in resource choice at the individual level. However, genetic groups within the colony varied in their tendency to collect pollen versus nectar, and in lability of response to changes in foraging stimuli. Our data suggest that within-colony genotypic diversity contributes to a resilient foraging response to environmental variation.     

Heat shock response in the central nervous system

The heat shock response is induced in nervous tissue in a variety of clinically significant experimental models including ischemic brain injury (stroke), trauma, thermal stress and status epilepticus. Excessive excitatory neurotransmission or the inability to metabolically support normal levels of excitatory neurotransmission may contribute to neuronal death in the nervous system in many of the same pathophysiologic circumstances. We demonstrated that in vitro glutamate-neurotransmitter induced excitotoxicity is attenuated by the prior induction of the heat shock response. A short thermal stress induced a pattern of protein synthesis characteristic of the highly conserved heat shock response and increased the expression of heat shock protein (HSP) mRNA. Protein synthesis was necessary for the neuroprotective effect. The study of the mechanisms of heat shock mediated protection may lead to important clues as to the basic mechanisms underlying the molecular actions of the HSP and the factors important for excitotoxic neuronal injury. The clinical relevance of these findings in vitro is suggested by experiments performed by others in vivo demonstrating that pretreatment of animals with a submaximal thermal or ischemis stress confers protection from a subsequent ischemic insult.     

Metabolism of endosulfan in the Indian honey bee

Summary Metabolism of topical endosulfan in the Indian honey bee,Apis cerana indica F., was studied. The two endosulfan isomers were found to be interconvertible in bees. The products of metabolism were endosulfan sulfate, diol, ether, hydroxyether, lactone and two unknown compounds. Endosulfan diol, hydroxyether and lactone were found to be conjugated in excreta of the treated bees.Acknowledgment. Authors are grateful to the department of Horticulture and Food Technology for providing gas chromatography facilities.     

Juvenile hormone,vitellogenin and haemocyte composition in winter worker honeybees (Apis mellifera)

Summary The haemolymph of winter bees (long life span) in contrast to that of summer bees (short life span) has a low juvenile hormone titre and a high protein titre. It is suggested that the winter bee physiology may be brought about by low JH-production.     

Cellular responses to mild heat stress

Since its discovery in 1962 by Ritossa, the heat shock response has been extensively studied by a number of investigators to understand the molecular mechanism underlying the cellular response to heat stress. The most well characterized heat shock response is induction of the heat shock proteins that function as molecular chaperones and exert cell cycle regulatory and anti-apoptotic activities. While most investigators have focused their studies on the toxic effects of heat stress in organisms such as severe heat stress-induced cell cycle arrest and apoptosis, the cellular response to fever-ranged mild heat stress has been rather underestimated. However, the cellular response to mild heat stress is likely to be more important in a physiological sense than that to severe heat stress because the body temperature of homeothermic animals increases by only 1–2°C during febrile diseases. Here we provide information that mild heat stress does have some beneficial role in organisms via positively regulating cell proliferation and differentiation, and immune response in mammalian cells.Received 14 May 2004; received after revision 2 August 2004; accepted 16 August 2004     

Thermal conductivity of wax comb and its effect on heat balance in colonial honey bees (Apis mellifera L.)

Summary Wax comb was found to have a thermal conductivity of 0.36×10^–3 cal/cm sec °C. At low air temperatures, honey bees,Apis mellifera L., form clusters inbetween the combs in their nests. The combs provide insulation and the bee behavior actually increases the insulating effectiveness of the combs. When they form a compact living layer over the wax comb, the conductivity can be reduced to 0.065×10^–3 cal/cm see °C. Some aspects of the role of the wax comb in heat balance are examined in this paper.     

Pollen-collecting by stingless bees on cacao flowers

Summary Stingless bees (Trigona jaty) routinely visit the flowers ofTheobroma cacao (Sterculiaceae) in the Atlantic lowlands of Costa Rica. The bees collect pollen and behave as pollen thieves in flowers well exposed to direct sunlight in cacao plantations, and avoid flowers in heavy shade. Pollination rates are maximized, however, in heavy shade due to the high abundance of the small-bodied pollinating midges (Ceratopogonidae and Cecidomyiidae) found in such places. Pollen-thieving by stingless bees, therefore, may only impact on fruit set in cacao trees in direct sunlight, with only minimal to no impact in areas of cacao where natural pollinator activity is high.This research was funded by grants from The American Cocoa Research Institute of the United States of America. I thank T. Noeske for statistical assistance.